Additional data related to the characterization of colonic Tregs isolated from F1-SSP mice and from natural SSP compared with non-SSP mice at 28 dpi. (A) Gating strategy used for sorting Tregs from colonic lamina propria of F1-uninfected and F1-SSP mice at 28 dpi. (B) Uniform Manifold Approximation and Projection (UMAP) plot showing the clustering of colonic Treg based on condition and combination (uninfected/SSP). (C) Barplot showing the contribution of each condition to the different Treg clusters. (D) Representative FACS plots, frequency, and numbers of T-bet⁺ Tregs isolated from the colonic lamina propria (cLP) of uninfected (black circle), non-SSP (dark-pink circle), and SSP (light-blue circle) mice at 28 dpi (n = 5–10). Results are representative of at least two independent experiments and presented as means ± SEM. One-way ANOVA followed by post hoc Tukey’s test was performed for multiple groups comparisons (****P < 0.0001; **P < 0.01; *P < 0.05; ns = not significant). (E) Representative flow cytometry histograms showing expression levels of the indicated markers in colonic Tregs isolated from of uninfected (black line), non-SSP (dark-pink line), and SSP (light-blue line) mice at 28 dpi. (F) Violin plots showing the expression levels of Il2ra, Pdcd1, Ctla4, Tnf, and Ifng among clusters 0 and 1, present in F1-SSP mice, versus clusters 2 and 3, mostly represented in uninfected F1 mice.